【作者】 刘培培；

【导师】 李会珍；

【作者基本信息】 中北大学 ， 生物化工， 2011， 硕士

【摘要】 本研究以紫苏的子叶和下胚轴为外植体进行组织培养,建立了紫苏的离体再生体系,获得了正常的再生植株。本实验运用对比试验的方法,研究了影响紫苏再生的因素,同时对紫苏的遗传转化体系进行了初步探索。其实验结果表明:1、紫苏子叶的最佳分化培养基为MS+BA 1.0mg/L+IAA 1.5 mg/L,再生率最高80.0%,平均每外植体长芽2.3个,下胚轴的最佳分化培养基为MS+BA 3.0mg/L+NAA 0.3 mg/L,再生率为70.9%,平均每外植体长芽2.5个。再生芽的生根培养基为1/2MS+1.0mg/L BA+0.3mg/L IAA,生根率达到100%,移栽成活率在85%以上。2、以11天苗龄的紫苏苗作为实验材料,白砂糖添加30g/L,2,4-D预培养后转入最佳分化培养基,子叶暗培养12天,下胚轴暗培养8天后,给予每天14h光照处理,可以较大幅度提高紫苏外植体的再生率。培养基中添加4mg/L AgNO3的处理可使子叶外植体再生频率提高5.4%,下胚轴外植体的再生频率提高9.4%。3、遗传转化研究表明,下胚轴抗性芽的筛选压力为40mg/L Kan,转化苗生根的筛选压力为35 mg/L Kan,研究发现培养基中添加15 mg/L Kan时,每外植体长芽7.6个,为对照组的3.3倍,较大地提高了每个外植体的再生芽数。农杆菌侵染后,选用羧苄青霉素作为抑菌剂,将外植体与农杆菌共培养2天,转接到附加羧苄青霉素400mg/L的培养基上进行抑菌处理,抑菌效果效果最好。更多还原

【Abstract】 Perilla cotyledon and hypocotyl were used as explants for tissue culture in this research. This study established vitro regeneration system for perilla and got normal regenerative plants. Comparative experimental methods were used to study the factors affecting the regeneration for perilla, while the perilla genetic transformation system were studied as a preliminary exploration,The conclusions were obtained as follows:1. Perilla cotyledons best differentiation medium was MS + BA 1.0mg/L + IAA 1.5 mg/L, the highest regeneration rate was 80.0%, every cotyledon germinated 2.3 buds. The perilla hypocotyl best differentiation medium was MS+BA 3.0mg/L + NAA 0.3 mg/L, the rate of regeneration was 70.9%, every cotyledon germinated 2.5 buds. Regeneration of shoots rooting medium was 1/2MS +1.0 mg/L BA +0.3 mg/L IAA, the rooting rate was up to 100% and the survival rate was more than 85%.2. 11-days aged perilla was used as experimental material, added sugar 30g/L. The perilla was transferred after precultured in 2,4-D, cotyledons precultured for 12 days in darkness and hypocotyl precultured for 8 days in darknessthen cultured at the condition of 14h/d light. Cultured in the medium added 4mg/L AgNO3, cotyledons explant regeneration rates improve 5.4% and hypocotyl explant regeneration rates improve 9.4%.3. Perilla hypocotyl selection pressure for regeneration was 40mg/L Kan, the rooting selective pressure was 35 mg/L Kan, The experment shows that every explant germinated 7.6 buds in the medium added Kan 15 mg/L. It is 3.3 times as control group, and greatly improve the number of regenerated shoots. After Agrobacterium infection, choose Carbenicillin as antibacterial agent, after two days co-culture, the explant transfered to the medium of 400 mg/L Car can get the best antimicrobial activity.更多还原